Universal assay cartridge and methods of use

This application is a Non-Provisional of and claims the benefit of priority of U.S. Provisional Application No. 63/217,672, filed Jul. 1, 2021, the entire contents of which are incorporated herein by reference.

The present invention relates generally to the field of biochemical analysis, and in particular to sample cartridges for analyzing a fluid sample.

The analysis of fluids such as clinical or environmental fluids generally involves a series of processing steps, which may include chemical, optical, electrical, mechanical, thermal, or acoustical processing of the fluid samples. Whether incorporated into a bench-top instrument, a disposable cartridge, or a combination of the two, such processing typically involves complex fluidic assemblies and processing algorithms.

Conventional systems for processing fluid samples employ a series of chambers each configured for subjecting the fluid sample to a specific processing step. As the fluid sample flows through the system sequentially from chamber to chamber, the fluid sample undergoes the processing steps according to a specific protocol. Because different protocols require different configurations, conventional systems employing such sequential processing arrangements are not versatile or easily adaptable to different protocols.

In recent years, there has been considerable development in the field of biological testing devices that facilitate manipulate a fluid sample within a sample cartridge to prepare the sample for biological testing by polymerase chain reaction (PCR). One notable development in this field is the GeneXpert sample cartridge by Cepheid. The configuration and operation of these types of cartridges can be further understood by referring to U.S. Pat. No. 6,374,684 entitled “Fluid Control and Processing System,” and U.S. Pat. No. 8,048,386 entitled “Fluid Processing and Control.” While these sample cartridges represent a considerable advancement in the start of the art when developed, as with any precision instrument, there are certain challenges in regard to performance and use of such systems and processes. Moreover, the precise requirements of different target types (e.g. bacterial or viral) typically necessitates the development of specialized devices and cartridges for each type or class of target, such that testing for a panel of differing targets associated with multiple suspected diseases or conditions, multiple samples must be obtained and multiple cartridges utilized, which quickly becomes costly, cumbersome and time-consuming.

Thus, there is a need for sample cartridges that overcome various challenges observed with regard to performance. There is further need for sample cartridges that provide greater versatility in performing assays for range of differing targets. There is further need for such devices that performs a wide range of sample processing steps in a robust and consistent manner and that are compatible with existing technologies to reduce costs and improve patient access.

The present invention pertains to sample cartridge devices and associated components, particularly sample cartridge devices capable of performing sample preparation for various differing types of targets within the same cartridge, as well as associated methods of use.

In one aspect, the invention provides a sample cartridge for separating a desired analyte from the sample and for holding the analyte for chemical reaction and optical detection. The invention also pertains to an instrument module that receives the cartridge for sample processing and operates the cartridge to perform sample preparation and analytical testing. The desired analyte is typically intracellular material (e.g., nucleic acid, proteins, carbohydrates, or lipids). In a preferred use, the analyte is nucleic acid which the cartridge separates from the fluid sample and holds for amplification (e.g., using PCR or an isothermal amplification method) and optical detection.

In another aspect, the invention pertains to a sample cartridge that utilizes a valve body platform that allows for detection of enveloped and free nucleic acid targets. In some embodiments, the valve body includes a sample processing region or lysing chamber that provides for heat, mechanical, and/or chemical lysis. This allows a single cartridge to provide lysing for a multitude of differing types of target, thus, can be considered a “universal assay cartridge.” In some embodiments, the sample cartridge can perform processing and detection of both bacterial targets requiring mechanical lysing and viral targets suited for chemical lysing. In some embodiments, the improved valve assembly provides for a sample cartridge capable of combined capture and detection of targets that do not require heat and/or mechanical lysis as well as targets that do require heat and/or mechanical lysis. In some embodiments, such valve assemblies are compatible with existing instrument modules that currently operate conventional sample cartridges directed to only one type of lysing.

In some embodiments, the valve assembly interfaces with the existing cartridge body such that operation of the cartridge by the instrument module is substantially the same or similar as a conventional sample cartridge. In some embodiments, the instrument module includes updates in modified operating instructions to perform workflows of sample preparation that perform multiple operations, such as chemical lysing, heat lysing, and mechanical lysing, of a single fluid sample with the same sample cartridge. In some embodiments, the instrument module reads or obtains the information regarding a panel of assays being performed, then operates according to a workflow that corresponds to one or all of heat lysing, mechanical lysing and chemical lysing depending on the assays being performed.

In another aspect, the invention pertains to a valve assembly that improves performance in regard to any of: consistency of fluid flow and filtering, distribution of forces, and distribution of in-fill (e.g. glass beads) for mechanical lysing. The valve assembly can include additional features that improve upon performance and functionality of the valve assembly as compared to conventional valves assemblies of sample cartridges.

In some embodiments, the valve assembly includes a valve body that interfaces with a valve cap to define an interior sample processing region or lysing chamber therebetween, the valve cap and valve body securing a filter therebetween, a fluid inlet in the valve cap and a fluid outlet in the valve body. In some embodiments, the cap includes a boss feature that interfaces with the valve body and is reduced in height, as compared to the conventional design, so as to accommodate a thicker filter material. In some embodiments, the fluid flow path through the inlets and outlet and lysing chamber or sample processing region have been improved to smooth transitions and eliminate any sharp angles to improve fluid flow therethrough and reduce residual buffer carryover.

In some embodiments, support features are added to the valve cap and valve body within the sample processing region or lysing chamber defined therebetween so as to improve in-fill ability and to reduce filter stress.

In some embodiments, the improvements include utilizing one or more protrusions in the cap adjacent an inlet port so as to increase clearance between any filter and the cap to improve fluid flow of sample and improve flow of in-fill, such as glass beads, for mechanical lysing. The one or more protrusions can include one or more posts on either or both sides of the inlet or outlet ports. In some embodiments, the posts are oval shaped with a major axis extending in a direction of flow. Experimental results using these post features improved yields of in-fill of glass beads within the sample processing region from 70% to 90%.

In other embodiments, the improvements include one or more protrusions or posts extending from the valve body adjacent an outlet so as to improve fluid flow across the filter region by maintaining a suitable gap between the filter and the valve body. This feature can reduce maximum pressure, for example by 5 psi, during assay testing and avoid clogging of the filter. In some embodiments, the posts are oval shaped with a major axis extending in a direction of flow. In some embodiments, the valve body includes a series of ridges extending in the direction of the fluid flow. In embodiments, having one or more posts near the outlet, the ridges extend only partly across the chamber, for example, about ¾ or less across the chamber and the posts are disposed between the series of ridges and the outlet. Experimental results showed that a valve body that included support posts adjacent the outlet reduced filter tears up to 10% by considerably reducing stresses in the filter.

In some embodiments, the valve assembly utilizes filters that are laser cut, which studies have shown to have reduced tears by up to 10%. Conventional approaches typically utilize mechanical cutting means, such as die cutting.

In a preferred embodiment, the cartridge has a sample port for introducing a sample into the cartridge, and a sample flow path extending from the sample port. The cartridge also has a lysing chamber in the sample flow path. The lysing chamber contains at least one filter for capturing cells or viruses from the sample as the sample flows through the lysing chamber. The lysing chamber is defined by at least one wall having an external surface for contacting the transducer to sonicate the lysing chamber. Beads may optionally be disposed in the lysing chamber for rupturing the cells or viruses as the chamber is sonicated. The cartridge can also include a waste chamber in fluid communication with the lysing chamber via the sample flow path for receiving the remaining sample fluid after the sample flows through the lysing chamber. The cartridge can further include a third chamber connected to the lysing chamber via an analyte flow path for receiving the analyte separated from the sample. The third chamber is preferably a reaction chamber for chemically reacting and optically detecting the analyte. The cartridge also includes at least one flow controller (e.g., valves) for directing the sample into the waste chamber after the sample flows through the lysing chamber and for directing the analyte separated from the sample into the third chamber. The design of the cartridge permits the efficient processing of large sample volumes to enable the accurate detection of low concentration analytes.

In some embodiments, the sample cartridge employs a rotary valve configuration that allows fluidic communication between a fluid processing region selectively with a plurality of chambers including, for example, a sample chamber, a waste chamber, a wash chamber, a lysis chamber, and a mastermix or reagent chamber. The fluid flow among the fluid processing region and the chambers is controlled by adjusting the position of the rotary valve. In this way, the metering and distribution of fluids in the apparatus can be varied depending on the specific protocol.

In accordance with some aspects of the present invention, a fluid control and processing system comprises a housing having a plurality of chambers, and a valve body including a first fluid processing region continuously coupled fluidicly with a fluid displacement region. The fluid displacement region is depressurizable to draw fluid into the fluid displacement region and pressurizable to expel fluid from the fluid displacement region. The valve body includes a plurality of external ports. The first fluid processing region is fluidicly coupled with at least two of the external ports. The fluid displacement region is fluidicly coupled with at least one of the external ports of the valve body. The valve body is adjustable with respect to the housing to allow the external ports to be placed selectively in fluidic communication with the plurality of chambers. At least one of the plurality of chambers is a processing chamber including at least one port for selectively communicating with at least one of the external ports of the valve body. The processing chamber provides an additional fluid processing region.

In some embodiments, at least one of the fluid processing regions in the valve body or in the processing chamber contains a fluid processing material which is an enrichment material or a depletion material. The fluid processing material may comprise at least one solid phase material. The solid phase material may comprise at least one of beads, fibers, membranes, filter paper, glass wool, polymers, cellulose fibers, and gels. In some embodiments, the filter is formed of glass fibers to facilitate affinity binding with the nucleic acids. In some embodiments, the filter has a nominal pore size of 0.2 to 2 um, preferably 0.5 to 1 um, typically about 0.7 um. In some embodiments, the cartridge includes glass beads for mechanical lysing, the glass beads having a nominal diameter of about 200 um or less, typically about 100 um. In some embodiments, the filter is a glass fiber disk without acrylic binder. In some embodiments, the filter material has a nominal thickness between 400 um and 450 um, typically about 420 um. In some embodiments, the cut filter is anywhere between 0.375″-0.400″ in diameter, with the nominal diameter being around 0.385″ or 9779 um. The fluid processing material may comprise a filter and beads, and in some embodiments comprises at least two types of beads. In some embodiments, a single type of solid phase material is used to perform at least two different functions which are selected from the group consisting of cell capture, cell lysis, binding of analyte, and binding of unwanted material. In some embodiments, the processing chamber includes a receiving area for receiving a processing module containing an enrichment material or a depletion material. In a specific embodiment, at least one of the chambers is a reagent chamber containing dried or lyophilized reagents. In some embodiments, the fluid processing material comprises at least one liquid phase material, such as ficoll, dextran, polyethylene glycol, and sucrose. The fluid processing material is contained in the fluid processing region by one or more frits. In a specific embodiment, the external ports are disposed on a generally planar external port surface of the valve body.

In some embodiments, the filter materials (e.g. glass beads, glass fibers) can be chemically treated to enhance performance. In some embodiment, the filter materials are chemically treated to improve binding and/or separation for isolation and purification of nucleic acids from nucleic-acid containing samples passed through the filter material. In some embodiments, the chemical treatment can include bonding of a compound to the filter material. In some embodiments, the compound comprises a DNA binding ligand, such as an amino-containing compound and can be used as a separating material for nucleic acid isolation. Particularly, the DNA binding ligand on the surface of the glass support provides high nucleic acid binding capacity for isolating the nucleic acid from a sample. In some embodiments, the compound is chemically bonded to the glass material via a linker, such as by an oligoethylene linker or a PEG oligomer. Suitable chemical treatments are described in U.S. Provisional Application No. 63/337,014 filed Apr. 29, 2022, entitled “Nucleic Acid Extraction and Isolation with Heat Labile Silanes and Chemically Modified Solid Supports,” the entire contents of which are incorporated herein by reference for all purposes. In some embodiments, the glass filter materials (e.g. glass fibers, beads) can be reacted with a silanizing group to obtain the separating materials disclosed herein. Accordingly, the silanol groups of the glass fibers can be reacted with compounds represented by the formula Y-(L)y-SiX3, wherein each X is independently selected from halogen, alkoxy, dialkylamino, trifluoromethanesulfonate, or a straight, branched, or cyclic alkyl; L is an optional linker such as an alkylene, heteroalkylene linker group, cyanuric chloride, an alkylamine, or a combination thereof and which may be optionally substituted; and Y is a DNA binding ligand, as described herein. The reaction of glass fibers with the compounds described herein provides in glass fibers surface DNA binding groups. The DNA binding ligand or the substituent Y can comprise a plurality of amine groups; a plurality of amide groups; or a combination thereof. For example, the DNA binding ligand or Y can comprise at least two, at least three, at least four, at least five, at least six amine or amide groups, or a combination thereof. In some embodiments, the DNA binding ligand or Y comprises an alkylamine group, an imidazole group, or a combination thereof. Representative examples of the amine group include spermine, methylamine, ethylamine, propylamine, ethylenediamine, diethylene triamine, 1,3-dimethyldipropyl-enediamine, 3-(2-aminoethyl)aminopropyl, (2-aminoethyl) trimethylammonium hydrochloride, tris(2-aminoethyl)amine, or a combination thereof. In some embodiments, the filter materials can comprise aminopropyl (AP) coated glass fiber filters (AP-GFF), glass beads, glass filter fibers, or other suitable solid support or fiber materials known to persons of skill in the art.

In accordance with another aspect of the invention, a fluid control and processing system comprises a housing having a plurality of chambers, and a valve body including a fluid processing region continuously coupled fluidicly with a fluid displacement region. The fluid displacement region is depressurizable to draw fluid into the fluid displacement region and pressurizable to expel fluid from the fluid displacement region. The valve body includes at least one external port, the fluid processing region is fluidicly coupled with at least one external port, and the fluid displacement region is fluidicly coupled with at least one external port of the valve body. The valve body is adjustable with respect to the housing to allow the at least one external port to be placed selectively in fluidic communication with the plurality of chambers.

In some embodiments, the sample cartridge employs a rotary valve configuration to control fluidic movement within the cartridge that allows for selective fluidic communication between a fluid sample processing region and a plurality of chambers in the cartridge. Non-limiting exemplary chambers can include, a sample chamber, a reagent chamber, a waste chamber, a wash chamber, a lysate chamber, an amplification chamber, and a reaction chamber. The fluid flow among the fluid sample processing region and the chambers is controlled by adjusting the position of the rotary valve. In this way, the metering and distribution of fluids in the cartridge can be varied depending on the specific protocol, which allows sample preparation to be adaptable to different protocols such as may be associated with a particular sample type for different types of analysis or different types of samples. For example, the sample cartridge can include a means for cell lysis, e.g., a sonication means so that bacteria and cells in a fluid sample to be analyzed can be lysed. Additional lysis means suitable for use with the instant invention are well known to persons of skill in the art, and can include, chemical lysis, mechanical lysis, and thermal lysis. In some embodiments, the sample includes bacteria, eukaryotic cells, prokaryotic cells, parasites, or viral particles.

In some embodiments, sample processing comprises sample processing steps that are performed from initial sample preparation steps, intermediate processing steps, and further processing steps to facilitate a detection of a target analyte in the biological sample with an attached reaction vessel. For example, sample processing can include preliminary preparation steps, such as filtering, grinding, mincing, concentrating, trapping debris or purifying a rough sample, or steps for fragmenting of DNA or RNA of the target analyte, such as by sonication or other mechanical or chemical means. Sample processing can include various intermediate processing steps, such as filtering, chromatography, or further processing of nucleic acids in the sample, including but not limited to chromatography, bisulfite treatment, reverse transcription, amplification, hybridization, ligation, or fragmentation of DNA or RNA. Sample processing may further include final processing steps, such as final amplification, hybridization, sequencing, chromatographic analysis, filtering and mixing with reagents for a reaction to detect the target analyte, which detection can include optical, chemical and/or electrical detection. While the sample cartridge typically performs analytical testing in an attached reaction tube or reaction vessel, it is appreciated that the sample cartridge can utilize various other means as well (e.g. semiconductor chip).

In some embodiments, the sample processing device can be a fluid control and processing system for controlling fluid flow among a plurality of chambers within a cartridge, the cartridge comprising a housing including a valve body having a fluid sample processing region continuously coupled fluidically with a fluid displacement chamber. The fluid displacement chamber is depressurizable to draw fluid into the fluid displacement chamber and pressurizable to expel fluid from the fluid displacement chamber. The fluid sample processing region includes a plurality of fluid transfer ports each fluidically coupled with one of a plurality of external ports of the valve body. The fluid displacement chamber is fluidically coupled with at least one of the external ports. The valve body is adjustable with respect to the plurality of chambers within the housing to allow the external ports to be placed selectively in fluidic communication with the plurality of chambers. In some embodiments, the valve body is adjustable with respect to the housing having multiple chambers, to place one external port at a time in fluidic communication with one of the chambers.

In some embodiments of the cartridge, the fluid sample processing region can be disposed between the fluid displacement chamber and at least one fluid transfer port. The term “fluid processing region” refers to a region in which a fluid sample is subject to processing including, without limitation, chemical, optical, electrical, mechanical, thermal, or acoustical processing. For example, chemical processing may include a chemical treatment, a change in pH, or an enzymatic treatment; optical processing may include exposure to UV or IR light; electrical processing may include electroporation, electrophoresis, or isoelectric focusing; mechanical processing may include mixing, filtering, pressurization, grinding or cell disruption; thermal processing may include heating or cooling from ambient temperature; and acoustical processing may include the use of ultrasound (e.g. ultrasonic lysis). In some embodiments, the fluid processing region may include an active member, such as a filter, to facilitate processing of the fluid. Additional active members suitable for use with the instant invention are well known to persons of skill in the art. In some embodiments, an energy transmitting member is operatively coupled with the fluid sample processing region for transmitting energy thereto to process fluid contained therein. In some embodiments, the valve body includes a crossover channel, and the valve body is adjustable with respect to the plurality of chambers to place the crossover channel in fluidic communication with two of the chambers concurrently.

The cartridge housing includes one or more branches that extend to one or more transfer ports to which a reaction vessel can be attached so as to facilitate transfer of fluid sample from a chamber of the cartridge into the reaction vessel. In some embodiments, the reaction vessel extends from the housing of the cartridge. These aspects can be understood further by referring to U.S. Pat. No. 8,048,386. It is understood that fluid may flow in either direction into or out of the transfer ports in various embodiments fluid flow is not limited in any particular direction. For example, in an embodiment having a pair of transfer ports, air may be pumped into or evacuated from one of the pair of transfer ports to facilitate flow of the fluid sample into a conduit of the reaction vessel through the fluid transfer port.

In some embodiments, methods for processing an unprepared sample can include steps of: receiving a sample cartridge in a cartridge receiver of a module, the sample cartridge including a biological fluid sample to be analyzed, a plurality of processing chambers fluidically interconnected by a moveable valve body; receiving an electronic instruction to process the biological sample into a prepared sample from the module; performing a sample preparation method in the sample cartridge to process the biological fluid sample into the prepared sample; transporting the prepared sample into a reaction vessel fluidically coupled with the sample cartridge; and performing analysis of the biological fluid sample within the reaction vessel. In some embodiments, transporting the sample may include steps of: moving a cartridge interface unit to move the valve body to change fluidic interconnections between the plurality of sample processing chambers; applying pressure to a pressure interface unit to move fluid between the plurality of processing chambers according to position of the valve body; and fluidically moving the prepared sample into the reaction vessel. Performing analysis of the fluid sample within the reaction vessel with the module. Any result of the analysis can be obtained by the module and communicated to various other devices as desired. In some embodiments, the sample cartridge can be coupled to various other diagnostic components, such as a silicon chip, or may transport the prepared fluid sample to other external diagnostic equipment.

The present invention relates generally to a system, device and methods for fluid sample manipulation and analysis, in particular, sample cartridges that facilitate processing and analytical testing of biological samples.

I. System Overview

In one aspect, the invention pertains to a sample cartridge that utilizes a valve body platform that allows for detection of enveloped and free nucleic acid targets. In some embodiments, the valve body includes a sample processing region or lysing chamber that provides for either or both mechanical and chemical lysis. This allows a single cartridge to provide lysing for a multitude of differing types of target, thus, can be considered a “universal assay cartridge.” In some embodiments, the sample cartridge can perform processing and detection of both bacterial targets requiring mechanical lysing and viral targets suited for chemical lysing.

The sample cartridge device can be any device configured to perform one or more process steps relating to preparation and/or analysis of a biological fluid sample according to any of the methods described herein. In some embodiments, the sample cartridge device is configured to perform at least sample preparation. The sample cartridge can further be configured to perform additional processes, such as detection of a target nucleic acid in a nucleic acid amplification test (NAAT), e.g., Polymerase Chain Reaction (PCR) assay, by use of a reaction tube attached to the sample cartridge. In some embodiments, the reaction tube extends from the body of the cartridge. Preparation of a fluid sample generally involves a series of processing steps, which can include chemical, electrical, mechanical, thermal, optical or acoustical processing steps according to a specific protocol. Such steps can be used to perform various sample preparation functions, such as cell capture, cell lysis, binding of analyte, and binding of unwanted material.

A sample cartridge suitable for use with the invention, includes one or more transfer ports through which the prepared fluid sample can be transported into an attached reaction vessel for analysis.illustrates an exemplary universal sample cartridgesuitable for sample preparation and analytics testing by PCR when received in an instrument module in accordance with some embodiments. The sample cartridge is attached with a reaction vessel(“PCR tube”) adapted for analysis of a fluid sample processed within the sample cartridge. In some embodiments the reaction tube extends form the cartridge body. Such a sample cartridgeincludes various components including a main housinghaving one or more chambers for processing of the fluid sample, which typically include sample preparation before analysis. The instrument module facilitates the processing steps needed to perform sample preparation and the prepared sample is transported through one of a pair of transfer ports into fluid conduit of the reaction vesselattached to the housing of the sample cartridge. The prepared biological fluid sample is then transported into a chamber of the reaction tubewhere the biological fluid sampleundergoes nucleic acid amplification. In some embodiments, the amplification is a polymerase chain reaction. In some embodiments, concurrent with the amplification of the biological fluid sample, an excitation means and an optical detection means of the module is used to detect optical emissions that indicate the presence or absence of a target nucleic acid analyte of interest, e.g., a bacteria, a virus, a pathogen, a toxin, or other target analyte. It is appreciated that such a reaction vessel could include various differing chambers, conduits, or micro-well arrays for use in detecting the target analyte. In some embodiments, the reaction tube can comprise multiple separate reaction chambers isolated from each other for the detection of different analytes. In some embodiments, the reaction tube can comprise 2, 3, 4, 5, 6, 7, 8, 9, 10 or more separate isolated reaction chambers. The sample cartridge can be provided with means to perform preparation of the biological fluid sample before transport into the reaction vessel. Any chemical reagent required for viral or cell lysis, or means for binding or detecting an analyte of interest (e.g. reagent beads) can be contained within one or more chambers of the sample cartridge, and as such can be used for sample preparation.

An exemplary use of a reaction vessel for analyzing a biological fluid sample is described in commonly assigned U.S. Pat. No. 6,818,185, entitled “Cartridge for Conducting a Chemical Reaction,” filed May 30, 2000, the entire contents of which are incorporated herein by reference for all purposes. Examples of the sample cartridge and associated module are shown and described in U.S. Pat. No. 6,374,684, entitled “Fluid Control and Processing System” filed Aug. 25, 2000, and U.S. Pat. No. 8,048,386, entitled “Fluid Processing and Control,” filed Feb. 25, 2002, incorporated herein by reference in their entirety for all purposes.

Various aspects of the sample cartridgecan be further understood by referring to U.S. Pat. No. 6,374,684, which described certain aspects of a sample cartridge in greater detail. Such sample cartridges can include a fluid control mechanism, such as a rotary fluid control valve, that is connected to the chambers of the sample cartridge. Rotation of the rotary fluid control valve permits fluidic communication between chambers and the valve so as to control flow of a biological fluid sample deposited in the cartridge into different chambers in which various reagents can be provided according to a particular protocol as needed to prepare the biological fluid sample for analysis. To operate the rotary valve, the cartridge processing module comprises a motor such as a stepper motor that is typically coupled to a drive train that engages with a feature of the valve in the sample cartridge to control movement of the valve in coordination with movement of the syringe, thereby resulting movement of the fluid sample according to the desired sample preparation protocol. The fluid metering and distribution function of the rotary valve according to a particular sample preparation protocol is demonstrated in U.S. Pat. No. 6,374,684.

II. Example Universal Assay Cartridge and Valve Assemblies

A. Overview

illustrates a universal sample cartridge devicefluidically coupled with a reaction vessel. The fluid sample cartridgeis adapted for insertion into a bay of a module having an instrument interface configured to perform one or more processing steps on a fluid sample contained within the fluid sample cartridge through manipulation of the fluid sample cartridge. The processing steps can include any steps associated with sample preparation, transport of fluid sample, and analytical testing. An instrument interface of the module is incorporated into the module within the bay in which cartridgeis received. At right, an exploded view of the sample cartridge assembly is shown, which includes the cartridge body, gasket, syringe, valve body, filter, valve capand cartridge base. While the basic function of these components may be similar to those of conventional sample cartridges, the valve assembly has been modified to considerably improve performance and to perform multiple types of sample preparation (e.g. mechanical and chemical lysing) with the same sample that would otherwise not be possible with conventional cartridges.

B. Examples Assays for Universal Assay Cartridges

In one aspect, the universal sample cartridge described herein can perform sample preparation and analytical testing for assays that are currently performed by conventional sample cartridges. For example, as shown in, one conventional sample cartridge (Cartridge A) is configured to perform analytical detections for targets including bacteria, spores, and hardy cells, which require mechanical lysis (e.g. ultrasonics, sonication) so that the released nucleic acids can be physically captured and detected. Another conventional sample cartridge (Cartridge C) is configured to perform analytical detections for target including virus, free DNA, and fragile cells, which can be easily lysed with chemical reagents to release the nucleic acids. Typically, these cartridges utilize nucleic acid binding. Currently, the capabilities of each sample configuration are such that each can only perform sample preparation for those designated targets. Advantageously, the universal sample cartridge described herein can perform both of these sample preparation tasks, individually, sequentially or in combination, such that the sample cartridge can replace the conventional cartridges as well as perform complicated assay panels that would otherwise not be achievable with a single sample cartridge.

For example, the universal sample cartridge can be used for the simultaneous detection of the major viral, parasitic and bacterial causes of undifferentiated febrile illness (UFI) in a Tropical Fever Assay panel, all of which can be performed by a sample cartridge utilizing the improved valve assembly described herein. Lysis requirements of possible target organisms responsible for UFI include both viral targets that require chemical lysis, parasitic and bacterial targets may require mechanical lysis. An example of such a test is shown in the table below.

Additional multi-target assay panels that can be developed for use with the universal sample cartridge may include a Gastrointestinal (GI) Panel, Breast Cancer Panel, and Bacterial Agents or any mixed-target panel. As shown, the differing targets within a single panel can include any of viral targets, fungal targets, parasitic targets, and bacterial targets, or any combination thereof.

C. Example Valve Assemblies

illustrates a valve assembly of a universal sample cartridge, in accordance with aspects of the invention, as compared to valve assemblies in conventional cartridge (A, B, C). Cartridge A performs only mechanical lysing for more hardy targets, and Cartridges B and C perform only chemical lysing for viruses, free NA or more fragile targets. In all such cartridges, the valve assembly includes the syringe tube, valve body, and valve cap. In one aspect, the additional capabilities of the valve assembly of the universal sample cartridge rely in part on the filter, features of the valve body and cap, as well as the particular workflow sequence performed by the instrument interface of the module.

In some embodiments, the filter is configured to accommodate glass beads to further facilitate mechanical lysis of hardy targets, as shown in. In this embodiment, the filteris formed of glass fibers and has a 0.7 um pore size. In contrast, Cartridge A utilizes a filter formed as a disk of a polymer film (i.e., PCTE), which while suitable for mechanical lysing, but not suited for chemical lysing. By utilizing a filter having a pore size of 0.7 um, the filter is suitable for receiving suitably sized glass beads for mechanical lysing. Utilizing glass fibers to form the filter facilitates affinity bonding with the free nucleic acid released by chemical lysing. Thus, this filter is suited for both mechanical and chemical lysing.

illustrate the valve cap, valve bodyand filterof a valve assembly of a universal sample cartridge, in accordance with some embodiments. The valve capand valve bodyinclude interlocking portions on their peripheries that engage with each so that interior circular regions interface to form a sample processing region or lysing chamber. The cap and valve body include interior circular regions,that interface to form an interior sample processing region or lysing chamber. The filteris sized to be secure between the valve cap and valve body

illustrate valve caps in accordance with some embodiments.shows a valve cap′ in accordance with some embodiments, andshows a valve capof a similar design that further includes a pair of posts. As shown, the valve caps include an interior circular regionthat defines the lysing chamber when interfaced with the valve body and a fluidic inletthrough which fluid sample and any glass bead infill enters the lysing chamber. In the design of, the valve cap further includes a pair of protrusions or posts, which presses the filter away from the inlet so as to improve consistency of glass infill across the filter so as to improve mechanical lysing.

illustrate valve bodies in accordance with some embodiments.shows a valve body′ in accordance with some embodiments, andshows a valve capof a similar design that further includes a pair of posts. As shown, the valve bodies includes the interior circular regionthat defines the lysing chamber when interfaced with the valve cap and a fluidic outletthrough which fluid sample exits the lysing chamber. In the design of, the valve body further includes a pair of protrusions or posts, which inhibits deflection of the filter toward the outlet so as to reduce pressure peaks on the filter and reduce tearing.

illustrate cross-sectional views of a valve body of a conventional valve assembly () and a valve body of an improved valve assembly () in accordance with some embodiments. As can be seen, the edges of the chamber or filter pocket have been smoothed such that sharp edges are eliminated, which reduces uneven flow and pressure distributions through the chamber and promote uniform flow through the lysing chamber.